2. Academic Validation
  3. In vitro functional characterization of novel nociceptin/orphanin FQ receptor agonists in recombinant and native preparations

In vitro functional characterization of novel nociceptin/orphanin FQ receptor agonists in recombinant and native preparations

  • Eur J Pharmacol. 2016 Dec 15:793:1-13. doi: 10.1016/j.ejphar.2016.10.025.
Federica Ferrari 1,  Maria Camilla Cerlesi 1,  Davide Malfacini 1,  Laila Asth 2,  Elaine C Gavioli 2,  Blair V Journigan 3,  Uma Gayathri Kamakolanu 3,  Michael E Meyer 3,  Dennis Yasuda 3,  Willma E Polgar 4,  Anna Rizzi 1,  Remo Guerrini 5,  Chiara Ruzza 1,  Nurulain T Zaveri 3,  Girolamo Calo 6
Affiliations

Affiliations

  • 1 Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
  • 2 Behavioral Pharmacology Laboratory, Department of Biophysics and Pharmacology, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
  • 3 Astraea Therapeutics, LLC. 320 Logue Avenue, Mountain View, CA, USA.
  • 4 SRI International, Biosciences Division, 333 Ravenswood Avenue, Menlo Park, CA, USA.
  • 5 Department of Chemical and Pharmaceutical Sciences and LTTA, University of Ferrara, Italy.
  • 6 Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy. Electronic address: [email protected].
PMID: 27780725 DOI: 10.1016/j.ejphar.2016.10.025
Abstract

Nociceptin/Orphanin FQ (N/OFQ) regulates several biological functions via selective activation of the N/OFQ receptor (NOP). In this study novel nonpeptide NOP ligands were characterized in vitro in receptor binding and [35S]GTPγS stimulated binding in membranes of cells expressing human NOP and classical opioid receptors, calcium mobilization assay in cells coexpressing the receptors and chimeric G proteins, bioluminescence resonance energy transfer (BRET) based assay for studying NOP receptor interaction with G protein and Arrestin, the electrically stimulated mouse vas deferens and the mouse colon bioassays. The action of the AT compounds were compared with standard NOP agonists (N/OFQ and Ro 65-6570) and the NOP selective antagonist SB-612111. AT compounds displayed high NOP affinity and behaved as NOP agonists in all the functional assays consistently showing the following rank order of potency AT-127≥AT-090≥AT-035>AT-004= AT-001. AT compounds behaved as NOP full agonists in the calcium mobilization and mouse colon assays and as partial agonists in the [35S]GTPγS and BRET assays. Interestingly AT-090 and AT-127, contrary to standard nonpeptide agonists that display G protein biased agonism, behaved as an unbiased agonists. AT-090 and AT-127 displayed higher NOP selectivity than Ro 65-6570 at native mouse receptors. AT-090 and AT-127 might be useful pharmacological tools for investigating the therapeutic potential of NOP partial agonists.

Keywords

Bioluminescence resonance energy transfer; Calcium mobilization and mouse colon vas deferens and assays; NOP and classical opioid receptors; Receptor and [(35)S]GTPγS binding.

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